A Modified DMAB-PAS Method

Abstract

A modified method of staining paraffin embedded tissue is described. The procedure is a combination of p-dimethylaminobenzaldehyde and periodic acid-Schiff (DMAB/PAS) techniques with a subbing solution or albumin instead of celloidin as the adhesive. Blocking reactions to demonstrate the specificity of the stains were performed with performic acid (DMAB) and diastase (PAS). The modified technique differentially stains tryptophan and mucopolysaccharides in stomach and pancreas tissue: the DMAB reaction sites are deep blue; the cytoplasm of the cells appears unstained, and the nuclei and basal membranes are colored magenta because of the PAS.

The combined method has the advantages of both the DMAB and PAS. It is simple to perform and requires only 15–20 minutes. This modified method is technically successful, produces permanent slides for further study, and may be adaptable to other tissues. (The J Histotechnol 13:209, 1990)     

A Modified Blood Stain,Useful in the Diagnosis and Evaluation of Hematologic Elements

Abstract

A modified blood stain has been developed for the demonstration and differentiation of cell types in peripheral blood or marrow smears, parasites in blood, and tissue imprint preparations. It is available as a powder extracted from known quantities of dyes of unpolychromed methylene blue, azure II and eosin Y. The powder keeps indefinitely without any appreciable loss of potency, and it is dissolved in 100% methanol or in equal parts of glycerol and methanol. The method of staining is simple and rapid, and the tinctorial patterns closely resemble those of the conventional blood stains, such as Giemsa, Wright, and Leishman. It is specific, stable, reproducible, and as the stain solution ages the staining quality improves.     

Staining efficacy assessment of a differential routine and special stains for pathological stromal calcifications in maxillofacial lesions

ABSTRACT

Head and neck connective tissue lesions may have diverse calcifications within the fibrous connective tissue stroma. The perplexity involved in the identification and determination of the nature or degree of calcification through routine hematoxylin and eosin (H&E) stains necessitates the usage of a specific, simple, and cost- and time-effective differential staining techniques. The aim of the present study was to develop criteria to distinguish bone formation from bone resorption using methylene blue-acid fuchsin (MB/AF) stain and the role of collagen fibers in the identification of stromal calcifications using polarizing microscopy with picrosirius red stain. Twenty cases with pathological diagnoses for various stromal calcifications in maxillofacial lesions were retrieved from the departmental archives. Decalcified formalin fixed paraffin embedded tissue sections were stained with hematoxylin and eosin, Masson’s trichrome (MT), methylene blue-acid fuchsin (MB/AF), and picrosirius red. The stained sections were assessed to identify the calcifications found in the surrounding connective tissue stroma. It was observed that most cases showed maximum staining intensity with MB/AF stain as compared to the other staining methods. Moreover, the results suggested that contrast between calcification and stromal soft tissue was best distinguished with the MB/AF stain except in the case of dystrophic calcifications. Along with this, polarizing microscopy with picrosirius red enables better characterization of stromal components. Although the H&E stain and a connective tissue stain i.e. Masson’s trichrome, are employed routinely in histopathology; the use of special stains such as MB-AF and picrosirius red facilitates the identification of calcifications from the stromal tissues.     

A Combination Stain for Acid Fast Bacilli and Fungi

Abstract

A combination stain for acid fast bacilli and fungi is described. The method is simple and it provides a reliable method of recognizing both organisms in tissue sections simultaneously in one setting. Basically it is a combination of Beamer's modification of Fites for acid fast bacilli and Gridley's stain for fungi.     

Identification of Pneumocystis carinii by Quick Hematoxy in and Eosin Smear

Abstract

Intraoperative identification of Pneumocystic carinii generally involves the use of special stains, which are time consuming and require expertise. The hematoxylin-eosin (H & E) stain readily demonstrates the organism in smears but not in tissue sections. The method described affords identification of these organisms within 5-10 min. A smear obtained from bisected lung biopsy tissue is immediately fixed with cytosol spray, dried for a few sec, and stained with H & E. When slides are examined with either high dry or oil immersion objectives, the smear shows foamy amorphous clusters of organisms, macrophages, and epithelial cells. The method stains both the cysts and the trophozoites, and the results correlate with those from special stains on tissue sections. It is rapid, simple to perform, low in cost, and easy to interpret. (The J Histotechnol 14:179, 1991)     

Volume 13, March 1990—December 1990

Abstract

A microwave Giemsa method for paraffin embedded tissue sections is reported. It is a modification of an earlier stain by Churukian and Listinsky, in which warm glycerol is used to dissolve the dyes and Triton X-100 is added to the final solution. The procedure can be completed within one hour and fifteen minutes and is consistent and reliable for demonstrating a variety of cells and microorganisms, like rickettsia and gram-negative bacteria. The stain can be easily prepared in the laboratory, does not require a microscope to control differentiation, and is as good, if not better, than commercially available Giemsa stains. Results are more uniform and consistent than those obtained with the widely used Wolbach method. (The J Histotechnol 18:319, 1995).     

Variation of the Holmes Method for Histologic Staining of Bone Canaliculi

Abstract

The Holmes method for soft tissue staining of nerve fibers was modified to produce excellent detail of the canalicular network in decalcified bone tissue. With simple changes in timing and the meticulous use of fresh reagents, we were able to stain bone lacunae and canaliculi in a variety of animal and human tissues. This technique produced more uniform and accurate results than the thionin/picric acid stains used previously for visualization of osteocytes. The morphologic results can be used for diagnostic purposes as well as stereological studies where the osteocyte network is of importance. (The J Histotechnol 16:355, 1993)     

A Modification of Kreyberg's Method for Demonstrating Keratin and Mucin

Abstract

A modification of Kreyberg's method for demonstrating keratin and acid mucopolysaccharides is described. The method substitutes alcian blue 8GX for alcian green 2GX and acidified metanil yellow in place of alcoholic saffron. Staining results with the method have been consistently satisfactory for demonstrating keratin and acid mucopolysaccharides in a variety of tumors.     

Modification of Brain Processing Techniques: Tissue Embedding in Glycol Methacrylate and Stain Modifications

Abstract

Increased interest in quantitation of the histopathological changes in a variety of neurological disorders (including neurodegenerative disease such as Alzheimer's disease) continues in an attempt to develop specific clinical-histopathological correlations. Most previous efforts at quantitation have used paraffin embedded sections of brain tissue, although plastic embedded sections have recently become a preferable alternative because they provide greatly reduced tissue shrinkage and distortion during processing, and greater clarity and improved resolution to the tissue sections. We have developed techniques for glycol methacrylate embedding and sectioning of brain tissue blocks on a standard histology laboratory microtome. In addition, we have modified routine diagnostic and investigational neurohistological stains for use in glycol methacrylate embedded brain sections, including hematoxylin and eosin, modified Bielschowsky stain, Jamarri silver technique, Einarson's Nissl stain, gallocyanin-Darrow red myelin stain, and the thioflavine-S-hematoxylin stain. The use of plastic embedded sections with appropriate stains will permit critical histopathological evaluation of nervous system tissue from patients with a variety of neurological disorders. (J Histotechnol 12:201, 1989)     

Enhancement of the Paraldehyde-Fuchsin Staining

Abstract

We have modified the aldehyde-fuchsin (PAF) stain to enhance its ability to simultaneous demonstration of different tissue compounds. We have substituted the light green dye of Hallni mixture with fast green dye and extended the reduction time by oxalic acid. The resulting stain provides better visualization of several tissue elements, such as the distinction between the nucleus and secretion products of goblet cells and between elastic and collagen fibers and others tissue elements. (The J Histotechnol 21:147, 1998)     

The Denn-Golgi stain — A Modified Defano's Silver Impregnation Stain for Dendritic Spines and Neurofibrils

Abstract

A modification of Defano's method for Golgi apparatus is described with emphasis on reduced overall stain time and better stain definition. This method utilizes a variation of widely used silver nitrate impregnation solution modified for a specific research need. The addition of a counterstain provides improved contrast and definition of dendritic spines in the Golgi apparatus of brain tissue. This modified method provides consistent, reproducible results for large numbers of brain specimens, and as such is believed to be superior to the Defano method.     

Citrate Buffer Alternative to Picric Acid for Masson Trichrome Stain

Abstract

An alternative to picric acid as a mordant in the staining of cytoplasm, keratin, muscle fibers, and intercellular fibers in the Masson trichrome stain is a citrate acid-sodium citrate buffer used as the microwave pretreatment of the tissue mounted on a slide. The Masson trichrome stain, a combination of Biebrich scarlet-acid fuchsin and aniline blue dye can be done within 1 hr and avoids the hazards associated with picric acid, a highly explosive compound in the dry state. (The J Histotechnol 19:341–342, 1996)     

NSH in Action

Abstract

A method for staining various structures of chondro — osseous trabeculae of the growth plate is described. A pseudo Schiff reagent made from crystal violet is used in the PAS sequence as the stain for calcified cartilage. Prestaining in saturated picric acid differentiated between calcified bone and osteoid tissue before staining with a modified Goldner's or Mallory's procedure. The method has been particularly suitable for histomorphometric studies of rat growth plates using an automatic numerized image analyzer system.     

A Light Resistant,Permanent Eosin Counterstain For Hematoxylin

Abstract

Many eosin dyes now sold stain cytoplasm in varying shades, often masking the tissue components under study. We use an eosin Y solution as a counterstain for consistent, permanent contrast among tissue components, even in the presence of light.     

A Rapid Method For Methyl Methacrylate Embedding of Mineralized Bone Specimens

Abstract

A method for methyl methacrylate embedding of mineralized bone specimens is described which takes only one week to complete. The resulting tissue block is easy to section and the four micron thick sections can be deplasticized and stained with routine or special stains.     

A Simple Device for Staining Electron Microscopy Grids

A simple holder for electron microscopy grids is described. It consists of a rod covered with dental wax (Ladd, Cat. No. 32165). Multiple slits for holding grids are made in the wax. The rod is attached to a rubber cork that fits into the test tubes filled with staining or rinsing solutions. The device has simplified and standardized the staining procedure, and stain precipitates due to air exposure are avoided.     

Commercially available fluorescein-conjugated monoclonal antibody for determining the in vitro activity of antimicrobial agents againstChlamydia trachomatis

A simple, reproducible method for determining the antibiotic susceptibility of chlamydial isolates is described. Minimum inhibitory and lethal concentrations (MICs and MLCs) were determined for tetracycline and erythromycin titrated against a recent clinical isolate of Chlamydia trachomatis in McCoy cell cultures. A fluorescent antibody stain was found to be more sensitive than giemsa staining, generally giving two-fold higher values for both MICs and MLCs.     

Identification of human megakaryocytes with rhodanile blue

Using the oxazine dye rhodanile blue, large typical megakaryocytes and small megakaryocytes (micromegakaryocytes) from the bone marrows of normal persons, and from patients with a variety of preleukemic disorders, acute lymphoblastic and nonlymphoblastic leukemia, chronic granulocytic leukemia, and idiopathic thrombocytopenic purpura as an example of nonmalignant but abnormal megakaryocytopoiesis, showed intense pink staining of the cytoplasm. This pink metachromasia was not obliterated by prior digestion with either diastase or ribonuclease, but was markedly diminished or obliterated by preincubation with hyaluronidase, suggesting that the stain may detect a high content of acid mucopolysaccharides in megakaryocytes. Since the stain is simple, direct, and reproducible, it may represent a useful addition to the cytochemistry of megakaryocytes and complement the more complex immunologic techniques available currently.     

Silver Labeling of Alzheimer Neurofibrillary Changes and Brain β-Amyloid

Abstract

Alzheimer neurofibrillary changes and brain β-amyloid can be stained selectively by silver impregnation on tissue sections and (in the case of neurofibrillary proteins) on sodium dodecyl sulfate-polyacrylamide gels. The stain can be applied to frozen sections of tissue from 10–150 μm thick, to similarly thick polyethylene glycol sections, or to 5–15 μm thick paraffin sections. The technique can also be applied to routinely fixed autopsy material. The technique takes advantage of the physical development of nucleation sites, thereby permitting tight control of the entire procedure. It is less expensive than immunocytochemical techniques, and facilitates processing of large numbers of sections through entire hemispheres of the human brain. (The J Histotechnol 16:335, 1993)     

Victoria Blue-Hematoxylin and Eosin Staining: A Useful Routine Stain for Demonstration of Venous Invasion by Cancer Cells

Abstract

Victoria blue (VB)-hematoxylin and eosin (H&E) staining, originally described by Yokokawa et al in 1983, has been used extensively in our surgical pathology lab as a routine stain for all surgical specimens with cancer. This simple staining sequence is useful for the demonstration of venous invasion by cancer cells as VB dye specifically demonstrates elastic fibers in the background of H&E-stained tissues. (The J Histotechnol 13:271, 1990)